The extracellular hydrolases of Mycobacterium Tuberculosis and their relation to obstructive pulmonary syndromes are to be studied. The specific aims are: 1) to study the mechanical properties of alveolar wall (rodent, man) before and after exposure to the extracellular products of M. tuberculosis, 2) to utilize changes in length-tension properties and in vitro assays to identify the active fractions while isolating the extracellular agent, 3) to characterize the isolated enzymes, and 4) ultimately to determine the site of action of these agents and the molecular effect. To isolate the agent(s) we plan to utilize the following techniques: 1) concentration using ammonium sulfate, ultrafiltration or freeze drying, 2) starch zone-electrophoresis or preparative gel electrophoresis, 3) affinity chromatography on protease inhibitor-Sepharose, elastin-Sepharose and collagen-Sepharose, and 4) gel filtration and ion-exchange chromatography. In an effort to determine the molecular effect of the enzymes, aliquots of lung parenchyma as well as aliquots of the various fractions of the parenchyma (collagen, elastin, proteoglycans) will be treated with the purified enzymes. The digests will be compared by gel-electrophoresis and the soluble fragment released by the treatment will be isolated and their composition determined. This is the first attempt we know of in which the product(s) of a specific bacterium (M. tuberculosis) associated with obstructive syndromes are to be identified. The identification and characterization of these agents will bring us closer to understanding the fundamental mechanism(s) which is responsible for the pathogenesis of obstructive syndromes (emphysema) as well as tuberculosis. Once these mechanisms are understood we may have a rational basis for the development of therapeutic inhibitors.